Ethanol Dependence and Stress Effects on Ethanol Drinking: CRF and Neurosteriods

描述(由申请人提供)：本研究建议是对美国国立卫生研究院关于恢复法资金可用于竞争性修订申请的公告(NOT-OD-09-058)的回应。目前为这项竞争性修订申请提供资金的父母基金(UOI AA014095)是NIAAA支持的INIA-Stress联盟的一个研究组成部分，该联盟专注于与过度饮酒行为有关的压力-乙醇相互作用。具体地说，该项目旨在研究与反复反复接触和戒酒有关的压力如何促进过量饮酒和增加与酒精依赖相关的复发风险的机制。利用酒精依赖和复发/过度饮酒的小鼠模型，重点研究CRF的作用。这项竞争性修订建议旨在扩大和加强我们的努力，以阐明与酒精依赖和饮酒行为升级相关的大脑奖励和应激途径中CRF活性的适应性变化。虽然父母赠款的研究集中在酒精依赖和反复饮酒的小鼠模型中CRF的神经适应性变化，但这些研究仅限于测量CRF本身的大脑区域mRNA变化(通过定量实时聚合酶链式反应(qRT-PCR)分析)和多肽含量(通过ELISA)。这项竞争性修订申请中提出的研究将扩展和补充与父母拨款有关的工作，重点是使用相同的酒精依赖和反复饮酒模型，研究cRFI和CRF2受体的转录和蛋白质变化。此外，除了通过qRT-PCR和免疫印迹分析脑CRF1和CRF2受体的mRNA和蛋白外，还将使用原位杂交技术更好地解析与酒精依赖和过度饮酒模型相关的CRF功能的神经适应性变化。总体而言，我们的目标是更全面地分析CRF应激神经肽系统在大脑奖赏和应激途径中功能的适应性变化，这些途径是与酒精依赖相关的应激引起的，并有助于过度饮酒行为和酒精中毒的发展。 与公共健康相关：虽然压力被视为酒精滥用和酒精中毒的一个重要因素，但压力和酒精饮酒行为之间的相互作用还没有被很好地理解。当人们在依赖的背景下考虑到压力对乙醇自我管理的作用/影响时，这一点尤其正确。这项研究项目的总体目标是更好地了解压力在促进过度饮酒和增加与酒精依赖相关的复发风险方面的作用。

DESCRIPTION (provided by applicant): This research proposal is submitted in response to the NIH announcement (NOT-OD-09-058) on availability of Recovery Act Funds for Competitive Revision Applications. The currently funded parent grant (UOI AA014095) to this competitive revision application is a research component of the NIAAA supported INIA- Stress Consortium, which is focused on stress-ethanol interactions in relation to excessive drinking behavior. Specifically, the project aims to examine mechanisms by which stress associated with repeated cycles of chronic ethanol exposure and withdrawal promotes excessive ethanol drinking and increased vulnerability to relapse associated with ethanol dependence. Utilizing a mouse model of ethanol dependence and relapse/excessive drinking, major emphasis is focused on studying the role of CRF. This competitive revision proposal is designed to extend and enhance our efforts to elucidate adaptive changes in CRF activity within brain reward and stress pathways that are associated with ethanol dependence and escalation of drinking behavior. While studies in the parent grant focus on neuroadaptive changes in CRF in the mouse model of ethanol dependence and relapse drinking, these studies are limited to measuring brain regional changes in mRNA (by quantitative real-time PCR (qRT-PCR) analysis) and peptide content (by ELISA) of CRF itself. Studies proposed in this competitive revision application will extend and complement work conducted in connection with the parent grant by focusing on transcriptional and protein changes in CRFI and CRF2 receptors using the same ethanol dependence and relapse drinking model. Further, in addition to analysis of brain CRF1 and CRF2 receptor mRNA and protein by qRT-PCR and immunoblotting, in situ hybridization procedures will be used to gain greater anatomical resolution of neuroadaptive changes in CRF function associated with the model of ethanol dependence and excessive drinking. Overall, the goal is to provide a more comprehensive analysis of adaptive changes in functioning of the CRF stress neuropeptide system within brain reward and stress pathways that result from stress associated with ethanol dependence and contribute to excessive drinking behavior and the development of alcoholism. PUBLIC HEALTH RELEVANCE: While stress has been viewed as an important contributing factor to alcohol abuse and alcoholism, the interaction between stress and ethanol drinking behavior is not well understood. This is especially true when one considers the role/impact of stress on ethanol self-administration in the context of dependence. The overall goal of this research project is to gain a better understanding about the role of stress in promoting excessive ethanol drinking and increased vulnerability to relapse associated with ethanol dependence.